Consideration on Efficient Recombinant Protein Production: Focus on Substrate Protein-Specific Compatibility Patterns of Molecular Chaperones.
Consideration on Efficient Recombinant Protein Production: Focus on Substrate Protein-Specific Compatibility Patterns of Molecular Chaperones.
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DOI：
10.1007/s10930-021-09995-4
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发表时间：
2021-10
期刊：
影响因子：
--
通讯作者：
Fedulov AV
中科院分区：
文献类型：
--
作者：
Yano N;Emi T;Gregory DJ;Fedulov AV
重组蛋白的表达有时需要分子伴侣的帮助，以有效地翻译后折叠成功能结构。然而，预测蛋白质底物与正确类型的伴侣蛋白的相容性以产生功能性蛋白质是一个令人生畏的问题。为了研究分子伴侣对具有不同特征的His标记的重组蛋白的影响的差异，我们使用过表达几个分子伴侣“团队”的大肠杆菌进行体外蛋白质表达：触发因子（TF），GroEL/GroES和DnaK/DnaJ/GrpE，单独或组合，目的是确定蛋白质二级结构是否可以作为分子伴侣成功的预测因子。蛋白A，其具有螺旋占优势的结构，表现出最有效的折叠与GroES/EL或TF伴侣单独，而蛋白B，其具有较少的螺旋结构，表现出显着的效果DnaK/J/GrpE系统单独。这种趋势也见于其他具有特殊性质的重组蛋白。在伴侣蛋白的辅助下，在22.5°C培养20小时比在37°C培养3小时更有效地合成两种蛋白质。这些研究结果提出了一种新的途径来研究伴侣与底物蛋白的相容性，并提出了生产功能蛋白的最佳培养条件，具有根据底物蛋白的性质预测分析伴侣成功的潜力。
Expression of recombinant proteins requires at times the aid of molecular chaperones for efficient post-translational folding into functional structure. However, predicting the compatibility of a protein substrate with the right type of chaperone to produce functional proteins is a daunting issue. To study the difference in effects of chaperones on His-tagged recombinant proteins with different characteristics, we performed in vitro proteins expression using Escherichia coli overexpressed with several chaperone ‘teams’: Trigger Factor (TF), GroEL/GroES and DnaK/DnaJ/GrpE, alone or in combinations, with the aim to determine whether protein secondary structure can serve as predictor for chaperone success. Protein A, which has a helix dominant structure, showed the most efficient folding with GroES/EL or TF chaperones alone, whereas Protein B, which has less helix in the structure, showed a remarkable effect on the DnaK/J/GrpE system alone. This tendency was also seen with other recombinant proteins with particular properties. With the chaperons’ assistance, both proteins were synthesized more efficiently in the culture at 22.5°C for 20 hours than at 37°C for 3 hours. These findings suggest a novel avenue to study compatibility of chaperones with substrate proteins and optimal culture conditions for producing functional proteins with a potential for predictive analysis of the success of chaperones based on the properties of the substrate protein.
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